The goal of this Phase 1 SBIR application is to identify and structurally characterize compounds in chocolate that could be used to treat tissue inflammation associated with gingivitis and periodontitis. Previous studies have demonstrated that water soluble extracts of cocoa bean husks and chocolate can contain activity that is either anti-inflammatory and/or dental plaque inhibiting. However, these extracts are practically unpalatable, a significant limitation for an oral formulation. Controlled evaluation of the structure-function relationships of potentially useful compounds arising during the chocolate manufacturing process has not been performed. In this proposal, extracts of characterized cocoa samples with known origin, fermentation profiles and specific heat treatment steps will be subjected to analytical characterization and preparative chromatographic separation and screened for the inhibition of Nf-kB activation in a mammalian cell line induced by lipopolysaccharides derived from Fusobacterium nucleatum. Fractions of chocolate extracts exhibiting Nf-kB antagonistic activity will be subject to initial characterization by mass spectrometry, and further separated to ensure that single chemical species are responsible for the observed activity. This Phase 1 research will lead to a Phase 2 project, where identified structures will be further characterized with respect to their mechanism of action and evaluated for their incorporation into a novel gum or mouthwash as a novel, safe, efficacious and cost-effective intervention for gingivitis and periodontitis. PUBLIC HEALTH RELEVANCE: Periodontal and gingival disease affects the majority of adults, can lead to tooth loss and has been associated with increased risks of cardiovascular disease and preterm birth, two conditions with high morbidity and mortality. There is compelling scientific evidence that extracts of cocoa display anti-inflammatory and dental plaque inhibiting characteristics but these chemistries are unpalatable for oral formulations. We propose a comprehensive evaluation of potentially useful compounds, with better flavor characteristics, generated during the chocolate manufacturing process that will lead to a better understanding of the therapeutic potential for cocoa in oral health applications and the development of products to prevent and control gingivitis and periodontitis.